猪带绦虫TSO45W-4BX与猪CD58分子在大肠埃希菌中的联合表达

目的 以猪CD58为分子佐剂，将CD58基因与猪带绦虫疫苗候选抗原基因TSO45W-4BX联合表达，寻找新型抗猪囊尾蚴疫苗。 方法 分别以重组质粒pGEM-4B和pGEM-CD58为模板，PCR扩增猪带绦虫TSO45W-4BX基因和猪CD58基因，将TSO45W-4BX与酶切处理的pGEX-4T-1定向连接，转化大肠埃希菌JM109，重组质粒经鉴定正确后，在其下游酶切插入CD58基因，PCR扩增和测序证明阅读框正确后，用异丙基-β-D-硫代半乳糖苷（IPTG）诱导表达，用十二烷基磺酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE））和蛋白质印迹法（Western blotting）分析表达产物的免疫活性。 结果 pGEX-4BX和pGEX-4BX/CD58分别表达Mr 41 000和Mr 69 000的融合蛋白，pGEX-4BX表达产物主要以可溶性形式存在，而pGEX-4BX/CD58却以包涵体形式存在，但两者都能被囊尾蚴病患者血清识别。 结论 TSO45W-4BX与CD58基因联合表达，TSO45W-4BX仍具有免疫活性。

Combined Expression of TSO45W-4BX from Taenia solium and Porcine CD58 in Escherichia coli

Objective To express the TSO45W-4BX of Taenia solium in combination with CD58 as a molecular adjuvant for improving the protective efficacy of the TSO45W-4BX recombinant vaccine. Methods TSO45W-4BX and porcine CD58 genes were amplified by PCR, using recombinant plasmids pGEM-4B and pGEM-CD58 as template respectively. The CD58 fragment was inserted into the recombinant plasmid pGEX-4T-1 with directly ligated TSO45W- 4BX, The transformant was induced with IPTG and followed by identifying the integrity of the recombinant containing TSO45W-4BX and porcine CD58 with PCR and sequencing. The products were analyzed by SDS-PAGE and Western blotting. Results The expression products of Mr69000 GST-4BX/CD58 and Mr41 000 GST-4BX were present mainly in the form of inclusion bodies and soluble substance respectively, and both were recognized by sera of cysticercosis patients. Conclusions The TSO45W-4BX co-expressed with porcine CD58 conserves its immune reactivity.